Systematic in vitro and in vivo characterization of Leukemia-inhibiting factor- and Fibroblast growth factor-derived porcine induced pluripotent stem cells

Jan O. Secher, Ahmet Ceylan, Gianluca Mazzoni, Kaveh Mashayekhi, Tong Li, Suchitra Muenthaisong, Troels T. Nielsen, Dong Li, Shengting Li, Stoyan Petkov, Susanna Cirera, Yonglun Luo, Lori Thombs, Haja N. Kadarmideen, A. Dinnyés, Lars Bolund, Bernard A J Roelen, Mette Schmidt, Henrik Callesen, Poul HyttelKristine K. Freude

Research output: Contribution to journalArticle

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Abstract

Derivation and stable maintenance of porcine induced pluripotent stem cells (piPSCs) is challenging. We herein systematically analyzed two piPSC lines, derived by lentiviral transduction and cultured under either leukemia inhibitory factor (LIF) or fibroblast growth factor (FGF) conditions, to shed more light on the underlying biological mechanisms of porcine pluripotency. LIF-derived piPSCs were more successful than their FGF-derived counterparts in the generation of in vitro chimeras and in teratoma formation. When LIF piPSCs chimeras were transferred into surrogate sows and allowed to develop, only their prescence within the embryonic membranes could be detected. Whole-transcriptome analysis of the piPSCs and porcine neonatal fibroblasts showed that they clustered together, but apart from the two pluripotent cell populations of early porcine embryos, indicating incomplete reprogramming. Indeed, bioinformatic analysis of the pluripotency-related gene network of the LIF- versus FGF-derived piPSCs revealed that ZFP42 (REX1) expression was absent in both piPSC-like cells, whereas it was expressed in the porcine inner cell mass at Day 7/8. A second striking difference was the expression of ATOH1 in piPSC-like cells, which was absent in the inner cell mass. Moreover, our gene expression analyses plus correlation analyses of known pluripotency genes identified unique relationships between pluripotency genes in the inner cell mass, which are to some extent, in the piPSC-like cells. This deficiency in downstream gene activation and divergent gene expression may be underlie the inability to derive germ line-transmitting piPSCs, and provides unique insight into which genes are necessary to achieve fully reprogrammed piPSCs. 84: 229–245, 2017.

Original languageEnglish
Pages (from-to)229-245
Number of pages17
JournalMolecular Reproduction and Development
Volume84
Issue number3
DOIs
Publication statusPublished - Mar 1 2017

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Induced Pluripotent Stem Cells
Fibroblast Growth Factors
Leukemia
Swine
Leukemia Inhibitory Factor
In Vitro Techniques
Genes
Gene Expression
Gene Regulatory Networks
Teratoma
Gene Expression Profiling
Computational Biology
Germ Cells
Transcriptional Activation
Embryonic Structures
Fibroblasts
Maintenance

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology
  • Cell Biology

Cite this

Systematic in vitro and in vivo characterization of Leukemia-inhibiting factor- and Fibroblast growth factor-derived porcine induced pluripotent stem cells. / Secher, Jan O.; Ceylan, Ahmet; Mazzoni, Gianluca; Mashayekhi, Kaveh; Li, Tong; Muenthaisong, Suchitra; Nielsen, Troels T.; Li, Dong; Li, Shengting; Petkov, Stoyan; Cirera, Susanna; Luo, Yonglun; Thombs, Lori; Kadarmideen, Haja N.; Dinnyés, A.; Bolund, Lars; Roelen, Bernard A J; Schmidt, Mette; Callesen, Henrik; Hyttel, Poul; Freude, Kristine K.

In: Molecular Reproduction and Development, Vol. 84, No. 3, 01.03.2017, p. 229-245.

Research output: Contribution to journalArticle

Secher, JO, Ceylan, A, Mazzoni, G, Mashayekhi, K, Li, T, Muenthaisong, S, Nielsen, TT, Li, D, Li, S, Petkov, S, Cirera, S, Luo, Y, Thombs, L, Kadarmideen, HN, Dinnyés, A, Bolund, L, Roelen, BAJ, Schmidt, M, Callesen, H, Hyttel, P & Freude, KK 2017, 'Systematic in vitro and in vivo characterization of Leukemia-inhibiting factor- and Fibroblast growth factor-derived porcine induced pluripotent stem cells', Molecular Reproduction and Development, vol. 84, no. 3, pp. 229-245. https://doi.org/10.1002/mrd.22771
Secher, Jan O. ; Ceylan, Ahmet ; Mazzoni, Gianluca ; Mashayekhi, Kaveh ; Li, Tong ; Muenthaisong, Suchitra ; Nielsen, Troels T. ; Li, Dong ; Li, Shengting ; Petkov, Stoyan ; Cirera, Susanna ; Luo, Yonglun ; Thombs, Lori ; Kadarmideen, Haja N. ; Dinnyés, A. ; Bolund, Lars ; Roelen, Bernard A J ; Schmidt, Mette ; Callesen, Henrik ; Hyttel, Poul ; Freude, Kristine K. / Systematic in vitro and in vivo characterization of Leukemia-inhibiting factor- and Fibroblast growth factor-derived porcine induced pluripotent stem cells. In: Molecular Reproduction and Development. 2017 ; Vol. 84, No. 3. pp. 229-245.
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AU - Mashayekhi, Kaveh

AU - Li, Tong

AU - Muenthaisong, Suchitra

AU - Nielsen, Troels T.

AU - Li, Dong

AU - Li, Shengting

AU - Petkov, Stoyan

AU - Cirera, Susanna

AU - Luo, Yonglun

AU - Thombs, Lori

AU - Kadarmideen, Haja N.

AU - Dinnyés, A.

AU - Bolund, Lars

AU - Roelen, Bernard A J

AU - Schmidt, Mette

AU - Callesen, Henrik

AU - Hyttel, Poul

AU - Freude, Kristine K.

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